LOCUS       sequence-327187-        6232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish kcnt1b gene CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6232)
  AUTHORS   Silic MR, Black MM, Zhang G
  TITLE     Phylogenetic and developmental analyses indicate complex functions 
            of Calcium-Activated Potassium Channels in zebrafish embryonic 
            development.
  JOURNAL   Dev Dyn. 2021 Mar 16. doi: 10.1002/dvdy.329.
  PUBMED    33728688
REFERENCE   2  (bases 1 to 6232)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dev Dyn. 
            2021 Mar 16. doi: 10.1002/dvdy.329."
COMMENT     Sequence Label: pENTR-D-TOPO-kcnt1b
FEATURES             Location/Qualifiers
     source          1..6232
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     119..136
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(295..322)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(414..500)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     549..571
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     563..580
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     564..580
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    596..695
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4384..4483)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4500..4519)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4501..4519)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4524..4540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4524..4540)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4642..4661)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4653..5462
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4720..4739)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5609..6197
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6098..6117
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
       61 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
      121 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
      181 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
      241 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
      301 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
      361 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
      421 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
      481 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
      541 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat
      601 aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt
      661 ttttataatg ccaactttgt acaaaaaagc aggctccgcg gccgccccct tcaccatgat
      721 gggaccgcgc agtcagtctc aggcagagcc aagcatgact ccgtcacgac gctcttccac
      781 tgctgctgac aggttgccta cggagactgt tcagagaaat aacagcagta atactggtgt
      841 tattctggat atttccacgc tgaaattggc agatgtggac tctgaggtcc ctcctctacc
      901 tccacgatac cgctttcgag atctgctgct tggggatcag tcatttcaga acgatgacag
      961 ggtgcaggtg gagttttacg taaacgaaaa cacatttaag gagaggttga agctcttctt
     1021 catcaaaaac caaaggtcaa gtctcagaat acgcctcttt aacctctcct taaagttgtt
     1081 aacatgcctg ctctacataa tcagagtctc acttgatgat ccaactaacc aggtcacatg
     1141 tgcacggaaa cactgcgcaa acatcacaga cgctctgtgc agcaaagcct gtgccaacta
     1201 tacagcaaac ctgaccgcag atcccacaca aataaactgg gagttaatat tttgggtgaa
     1261 tagaagagtt cctgtatggg cgatacaggt gacagtggcc ttaattagct tcttgcaagc
     1321 gatgctcctg gtgtatctca gctacaaggg caacatttgg gaacagattt tccaaatctc
     1381 cttcatcatt gagatgatca atacagtgcc attcataatt acaatctttt ggcctccgct
     1441 aaggaacata tttgttcctg tatttcttaa ctgttggctt gccaaatcag cgctggagaa
     1501 tatgattaat gatctccatc gtgccatcca gcgcacacac tctgccatgt tcaaccaggt
     1561 cctcatcctg atctgtacac tgctctgtct ggtcttcacc ggggcttgtg ggattcagca
     1621 cctggagcga gcagggaaga atctgacact ctttgacgct ttatatttct gcatcgtcac
     1681 cttctccact gttggatatg gagatgtgac gcccagaatc tggccctctc agctgctggt
     1741 ggtcatcatg atctgtgtgg cgctggtggt gctgccatta cagtttgagg agttggcgta
     1801 tctatggatg gagaggcaga agtcaggtgg aaactacagc agacaccgcg ctcagacaga
     1861 gaaacacgtg gtgctctgcg tcacctcact caaaatagac ctgctcatgg atttcctcaa
     1921 tgagttctat gcccacccga gactacagga ctattatgtg gtgatcctct gtcccacgga
     1981 gatggacatc caggtgcggc gaatactgca gatcccactg tggtctcagc gggtcatcta
     2041 cctccaaggc tctgcactga aggaccaaga cctcatgagg gccaagatgg atgatgcgga
     2101 ggcctgcttc atcctcagca gcagaaatga agtggaccgc accgctgctg atcatcagac
     2161 cattctgcga gcctgggctg tgaaggattt tgctccaaac tgtcctctat atgtccagat
     2221 tctcaaaccc gagaacaaat ttcatgtcaa atttgcagat cacgtggtgt gtgaagagga
     2281 gttcaagtac gccatgctgg ccctgaactg tgtgtgtccc gccacctcca cactggtcac
     2341 cctgctggtg cacacgtctc gaggacaaga aggtcagacg tctccagaac agtggcagag
     2401 gacttacggt cgctgctcgg ggaatgaagt ttatcacatc cgtttgatgg acagcaagtt
     2461 ttttggagaa tatgatggga agagtttcac ttatgcctcc ttccacgcac acaagaagta
     2521 tggtgtgtgt ctaatcggca caaagagaga agacaacaag agcattctcc tgaaccccgg
     2581 ccctcgccac atcatgtccg ccacagacac ctgctactac atcaacatca ccaaagagga
     2641 gaactcggcc ttcatcttca agcaggagga gaagcacagt aaaggcttat cagccgccgg
     2701 cgtgtacgac gctccgtctc gcctcccagt gcaaagcatc attgccagca tgggaactgt
     2761 ggccattgac ctgcagaacc cggatccacc ggctgacagc aataaactgg ctctgcccac
     2821 ggagaacgga gcgggcagcc gcagacccag cattgcccct gtgctggaga tcgcagactc
     2881 ctctgccatc ctgccctgtg atctcctcaa cgacccctca gaagacgaga ccaaccaatc
     2941 agatgaggat ggagcgccgg ccatagactg tgttaaaggc tatcctccaa actcccctta
     3001 cattggaagc tctccaacac tttgccacct tcttcaacag aaggcaccgt tttgctgtct
     3061 acgacttgac aagggttgtc gacacaatag tttcgaagac gccaaggcgt atggtttcaa
     3121 gaacaagttg attatcgttt ctgctgagac agcgggaaat ggcttgtaca acttcatagt
     3181 gccgttacgt gcctattacc gaccgcggaa ggagctgaat cccattgtgc tgctgctgga
     3241 taatccgcct gataatcatt tcctagaggc catctgctgt ttccctatgg tgtactacat
     3301 ggcgggaacc atagacaatt tggataacct gctgcagtgt ggcatcatct acgctgataa
     3361 cctggtggtg gtggataagg agagcactat gagcgcagag gaagactaca tggcggacgc
     3421 taaaaccata gtcaatgtgc agaccatgtt caggttgttt cctagtctca gcataatcac
     3481 tgagctaact catccgtcta atatgaggtt catgcagttc cgtgccaaag actgctactc
     3541 cctggctctc tccaaactcg aaaagataga gcgagataag ggatcgaatc tggcgttcat
     3601 gttccgcctg ccgttcgctg caggcagagt cttcagtatc agcatgctgg atactctgct
     3661 ttaccagtcg tttgtgaagg actatatgat tctcattgcg cggctcctcc tgggtttaga
     3721 caccaccccg ggctccggat acctctgtgc tatgaaggta acagaagagg acttatggat
     3781 tcgaacgtac gggagactct tccagaagct gtgttcctcc agcgcagaga ttcccatcgg
     3841 gatttaccgc accgaatccc acatgttctc ctcttcagag tctcaggtgt ctgtggagga
     3901 ttgcgaggac cagcgggacc acagggaatc ctggaaagag aaaactgccc ataggaactc
     3961 cacagggagc gaccaatcag agcatccgct cctgcgcaag aaaagcatgc agtgggcgag
     4021 acgcctaagc agaaaaaaca ccaaacctcc cagtcgagct gaacgcatca atcaacagcg
     4081 cctgaacctg taccggcgct ccgagaggca ggagctgtca gagctggtga agaaccgcat
     4141 gaagcattta ggcctgccca ctgtgggata cgatgaaatg aatgacaatc agaacacgtt
     4201 gtcatatgtg ctgataaacc ctccgccgga caccattctg gagctcaatg acattgtcta
     4261 catcatccgt tctgatccac tggcacatgt ccctgaaaat ctcccagggg ctccagctgg
     4321 agcgaagata cgccaagact atcggcccga gacaagggat gagacgcaag ggtgggcgcg
     4381 ccgacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
     4441 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc
     4501 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
     4561 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
     4621 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga
     4681 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     4741 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
     4801 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     4861 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     4921 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag
     4981 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     5041 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     5101 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     5161 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     5221 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     5281 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     5341 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     5401 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     5461 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     5521 gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     5581 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5641 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5701 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5761 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5821 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5881 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5941 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6001 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6061 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6121 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6181 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gc
//
